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Team:Warsaw/RBSmeasuremen/Results
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<div align="justify">As you can see below there are significant differences in RBS strength relative to B0034 between various fluorescent proteins. This is probably caused by influence of protein coding sequence on mRNA fold so different proteins will always have different expression levels from the same RBS and promoter. Differences reach 60% so you cannot predict expression level of your favorite protein basing on measurements with GFP.<br /> | <div align="justify">As you can see below there are significant differences in RBS strength relative to B0034 between various fluorescent proteins. This is probably caused by influence of protein coding sequence on mRNA fold so different proteins will always have different expression levels from the same RBS and promoter. Differences reach 60% so you cannot predict expression level of your favorite protein basing on measurements with GFP.<br /> | ||
In order to ensure that every protein will behave the same way with given expression driving part we need new approach. We need to deal with mRNA fold so we also need to get rid of different protein beginnings. One of solutions are our <a href="https://2011.igem.org/Team:Warsaw/ExpressionAdaptors">expression adapters.</a> | In order to ensure that every protein will behave the same way with given expression driving part we need new approach. We need to deal with mRNA fold so we also need to get rid of different protein beginnings. One of solutions are our <a href="https://2011.igem.org/Team:Warsaw/ExpressionAdaptors">expression adapters.</a> | ||
| - | <br> | + | <br><br> |
| - | The other | + | The other approach would be to improve expression strength prediction basing on existing measurements.<ul> |
| - | <li>It would require | + | <li>It would require creation of a database of measurements of each RBS with various proteins.</li> |
<li> The protein N terminal parts (e.g. first 10 aa) could be compared using blast against database of N-terminal sequences of proteins measured with particular RBS. </li> | <li> The protein N terminal parts (e.g. first 10 aa) could be compared using blast against database of N-terminal sequences of proteins measured with particular RBS. </li> | ||
<li>The best blast hit could be used to predict expression strength along with computational supporting tools e.g. RBS designer or RBS calculator </li></ul> | <li>The best blast hit could be used to predict expression strength along with computational supporting tools e.g. RBS designer or RBS calculator </li></ul> | ||
Unfortunately creating such database is currently not possible during summer project. So far we could tray predicting expression of proteins with similar N-terminal sequence to measured 5 fluorescent proteins. | Unfortunately creating such database is currently not possible during summer project. So far we could tray predicting expression of proteins with similar N-terminal sequence to measured 5 fluorescent proteins. | ||
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| + | Anyway, have a look at our results! | ||
| + | <br> | ||
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<div class="note">XFP expression driven by different RBS parts in numbers</div><br /> | <div class="note">XFP expression driven by different RBS parts in numbers</div><br /> | ||
<div align="center"><img src="https://static.igem.org/mediawiki/2011/f/f2/Tabelka.png" alt="ERROR" /></div><br /> | <div align="center"><img src="https://static.igem.org/mediawiki/2011/f/f2/Tabelka.png" alt="ERROR" /></div><br /> | ||
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Latest revision as of 22:34, 21 September 2011
RBS Measurement Results
As you can see below there are significant differences in RBS strength relative to B0034 between various fluorescent proteins. This is probably caused by influence of protein coding sequence on mRNA fold so different proteins will always have different expression levels from the same RBS and promoter. Differences reach 60% so you cannot predict expression level of your favorite protein basing on measurements with GFP.
In order to ensure that every protein will behave the same way with given expression driving part we need new approach. We need to deal with mRNA fold so we also need to get rid of different protein beginnings. One of solutions are our expression adapters.
The other approach would be to improve expression strength prediction basing on existing measurements.
Anyway, have a look at our results!
In order to ensure that every protein will behave the same way with given expression driving part we need new approach. We need to deal with mRNA fold so we also need to get rid of different protein beginnings. One of solutions are our expression adapters.
The other approach would be to improve expression strength prediction basing on existing measurements.
- It would require creation of a database of measurements of each RBS with various proteins.
- The protein N terminal parts (e.g. first 10 aa) could be compared using blast against database of N-terminal sequences of proteins measured with particular RBS.
- The best blast hit could be used to predict expression strength along with computational supporting tools e.g. RBS designer or RBS calculator
Anyway, have a look at our results!
Graphical representation of measurement results
XFP expression driven by different RBS parts in numbers
